Reliable LC-MS quantitative glycomics using iGlycoMab stable isotope labeled glycans as internal standards

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• 作者：Shiyue Zhou ; Nadia Tello ; Alex Harvey ; Barry Boyes ; Ron Orlando and Yehia Mechref
• 刊名：ELECTROPHORESIS
• 出版年：2016
• 出版时间：June 2016
• 年：2016
• 卷：37
• 期：11
• 页码：1489-1497
• 全文大小：1434K
• ISSN：1522-2683

文摘

Glycans have numerous functions in various biological processes and participate in the progress of diseases. Reliable quantitative glycomic profiling techniques could contribute to the understanding of the biological functions of glycans, and lead to the discovery of potential glycan biomarkers for diseases. Although LC-MS is a powerful analytical tool for quantitative glycomics, the variation of ionization efficiency and MS intensity bias are influencing quantitation reliability. Internal standards can be utilized for glycomic quantitation by MS-based methods to reduce variability. In this study, we used stable isotope labeled IgG2b monoclonal antibody, iGlycoMab, as an internal standard to reduce potential for errors and to reduce variabililty due to sample digestion, derivatization, and fluctuation of nanoESI efficiency in the LC-MS analysis of permethylated N-glycans released from model glycoproteins, human blood serum, and breast cancer cell line. We observed an unanticipated degradation of isotope labeled glycans, tracked a source of such degradation, and optimized a sample preparation protocol to minimize degradation of the internal standard glycans. All results indicated the effectiveness of using iGlycoMab to minimize errors originating from sample handling and instruments.